PCR of episomal plasmids

ALASTAIR MACKAY amackay at welchlink.welch.jhu.edu
Fri Jun 3 23:42:05 EST 1994


I agree with Brian Foley that the answer is PCR of whole cells.  It seems 
to me that a combination of positive controls and optimization has a good 
chance of working.  Positive controls:  x pg or fg of plasmid doping the 
whole cells, compared to this amount in absence of cells.  Optimization:  
Pay attention to Mg+2, do a checkerboard as Current Protocols suggests.  
Also check annealing temperature.  Consider adding TMAC (tetramethyl 
ammonium chloride) in the 10E-5 to 10E-6 molar range, I found it can help
increase specificity.
Good luck, Alastair



More information about the Methods mailing list